title: Genetic tailoring of N-linked oligosaccharides: the role of glucose residues in glycoprotein processing of Saccharomyces cerevisiae in vivo.
abstract: In higher eukaryotes a quality control system monitoring the folding state of glycoproteins is located in the ER and is composed of the proteins calnexin, calreticulin, glucosidase II, and UDP-glucose: glycoprotein glucosyltransferase. It is believed that the innermost glucose residue of the N- linked oligosaccharide of a glycoprotein serves as a tag in this control system and therefore performs an important function in the protein folding pathway. To address this function, we constructed Saccharomyces cerevisiae strains which contain nonglucosylated (G0), monoglucosylated (G1), or diglucosylated (G2) glycoproteins in the ER and used these strains to study the role of glucose residues in the ER processing of glycoproteins. These alterations of the oligosaccharide structure did not result in a growth phenotype, but the induction of the unfolded protein response upon treatment with DTT was much higher in G0 and G2 strains as compared to wild-type and G1 strains. Our results provide in vivo evidence that the G1 oligosaccharide is an active oligosaccharide structure in the ER glycoprotein processing pathway of S.cerevisiae. Furthermore, by analyzing N- linked oligosaccharides of the constructed strains we can directly show that no general glycoprotein glucosyltransferase exists in S. cerevisiae.
authors.affiliation: Division of Cell and Molecular Pathology, Institute for Microbiology, Zurich, Switzerland.
authors.forename: C A
authors.initials: CA
authors.lastname: Jakob
authors.forename: P
authors.initials: P
authors.lastname: Burda
authors.forename: S
authors.initials: S
authors.lastname: te Heesen
authors.forename: M
authors.initials: M
authors.lastname: Aebi
authors.forename: J
authors.initials: J
authors.lastname: Roth
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002236:Carbohydrate Conformation; D002268:Carboxypeptidases / Q000378:metabolism; D043402:Cathepsin A; D003433:Crosses, Genetic; D004229:Dithiothreitol / Q000494:pharmacology; D004789:Enzyme Activation; D005656:Fungal Proteins / Q000032:analysis; D005947:Glucose / Q000378:metabolism / Q000502:physiology*; D005964:Glucosyltransferases / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000201:enzymology; D008214:Lymphocytes / Q000201:enzymology; D009844:Oligosaccharides / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D000520:alpha-Glucosidases / Q000032:analysis
chemical_list: D005656:Fungal Proteins; D006023:Glycoproteins; D009844:Oligosaccharides; D005964:Glucosyltransferases; C072423:mannosylglycoprotein 1,3-glucosyltransferase; C034251:4-nitrophenyl-alpha-glucosidase; D000520:alpha-Glucosidases; D002268:Carboxypeptidases; C468629:CTSA protein, human; D043402:Cathepsin A; D005947:Glucose; D004229:Dithiothreitol